Abonnenc M, Pesse B, Tissot J-D, Barelli S, Lion N
Laboratoire R&D Produits, Transfusion Interrégionale CRS, Epalinges, Switzerland.
Vox Sang. 2017 May;112(4):367-378. doi: 10.1111/vox.12503. Epub 2017 Mar 23.
Autologous haematopoietic progenitor cell (HPC) is a prerequisite for high-dose chemotherapy in treatment of several haematologic and non-haematologic malignancies. HPCs are collected by apheresis and cryopreserved until infusion. Postinfusion adverse events have been in part related to the dimethyl sulphoxide (DMSO) used as cryoprotectant. The aim was to evaluate (i) an automated sequential washing procedure for DMSO removal in thawed HPC grafts and (ii) washing solutions in replacement of hydroxyethyl starch (HES)-based solutions.
A total of 26 HPC bags cryopreserved with 10% DMSO and intended for disposal were used. The Sepax-2 (Biosafe, Eysins, Switzerland) was evaluated using a sequential washing procedure. Outcomes were CD34 cell recovery and viability after washing.
The Ringer lactate supplemented or not with albumin 2·5-5% presented satisfactory results compared with HES solution in terms of CD34 cell recovery and viability after washing. However, the apparition of aggregates led us to renounce to these alternative solutions. Using HES solution and a sequential washing of three bags, we showed the elimination of 95% of the DMSO, a postwash viable CD34 cell recovery of 79·9 ± 9·4% and a total nucleated cell viability of 66·5 ± 9·3%.
The preclinical evaluation of an automated sequential washing procedure for DMSO removal in thawed HPC grafts has proven to be effective and comparable to previously published data. Despite our attempt to find an alternative solution to the HES solution, more efforts should be done on this side to reach a consensus on cryopreservation protocols.
自体造血祖细胞（HPC）是治疗多种血液系统和非血液系统恶性肿瘤时进行大剂量化疗的前提条件。HPC通过血细胞分离术采集并冷冻保存直至输注。输注后不良事件部分与用作冷冻保护剂的二甲基亚砜（DMSO）有关。本研究旨在评估（i）一种用于去除解冻后HPC移植物中DMSO的自动化顺序洗涤程序，以及（ii）替代基于羟乙基淀粉（HES）溶液的洗涤溶液。
共使用了26个用10%DMSO冷冻保存且拟丢弃的HPC袋。使用顺序洗涤程序对Sepax-2（Biosafe，瑞士艾辛斯）进行评估。观察指标为洗涤后CD34细胞回收率和活力。
与HES溶液相比，添加或不添加2.5 - 5%白蛋白的乳酸林格液在洗涤后CD34细胞回收率和活力方面表现出令人满意的结果。然而，聚集体的出现使我们放弃了这些替代溶液。使用HES溶液并对三个袋子进行顺序洗涤，我们发现可去除95%的DMSO，洗涤后活CD34细胞回收率为79.9±9.4%，总核细胞活力为66.5±9.3%。
对解冻后HPC移植物中DMSO去除的自动化顺序洗涤程序的临床前评估已证明是有效的，且与先前发表的数据相当。尽管我们试图寻找HES溶液的替代溶液，但在这方面仍需做出更多努力，以就冷冻保存方案达成共识。
